杜小正;袁博;张星华;田亮;张婷卓;李兴兰;张枫帆;王金海;
DU Xiao-zheng;YUAN Bo;ZHANG Xing-hua;TIAN Liang;ZHANG Ting-zhuo;LI Xing-lan;ZHANG Feng-fan;WANG Jin-hai;School of Acu-moxibustion and Massage,Gansu University of Chinese Medicine;Department of Acu-moxibustion,Gansu Provincial Hospital of Traditional Chinese Medicine;Department of Chinese Medicine,the Second Hospital of Lanzhou University;

• 1:甘肃中医药大学针灸推拿学院
• 2:甘肃省中医院针灸科
• 3:兰州大学第二医院中医科

摘要(Abstract)：

目的:采用代谢组学技术探讨热补针法治疗类风湿关节炎(RA)寒证的特异性作用机制。方法:青紫蓝家兔随机分为正常组、模型组、平补平泻组、捻转补法组、热补针法组,每组8只。以卵蛋白诱导联合低温冷冻法建立RA寒证模型。各治疗组分别运用平补平泻法、捻转补法、热补针法于双侧"足三里"穴进行针刺干预,治疗30min,每日1次,共治疗7d。治疗后观察各组家兔的痛阈值以及关节表面温度,采用气相色谱-质谱(GC-MS)检测结合模式识别法分析家兔血清中内源性代谢物的变化。结果:模型组家兔痛阈、关节表面温度较正常组降低(P<0.05),各针刺组干预后痛阈、关节表面温度较模型组升高(P<0.05),热补针法组高于平补平泻组、捻转补法组(P<0.05)。与正常组相比,模型组家兔血液代谢物中亮氨酸、缬氨酸、异亮氨酸、花生四烯酸的含量升高(P<0.05),α-酮戊二酸、柠檬酸、琥珀酸、葡萄糖、纤维糖、棕榈酸、硬脂酸、乳糖、d-核糖、D-甘露糖的含量降低(P<0.05)。与模型组比较,各针刺组血清中代谢物均出现不同程度的回调(P<0.05),且热补针法组对α-酮戊二酸、柠檬酸、琥珀酸、葡萄糖、纤维糖、d-核糖、D-甘露糖的回调明显优于平补平泻组和捻转补法组(P<0.05)。结论:热补针法治疗RA具有确切的疗效,可能与对三羧酸循环与糖代谢的相对特异性调控有关。
Objective To explore the specific mechanism of heat-reinforcing needling(HRN)for rheumatoid arthritis(RA)rabbits with cold syndrome based on metabolomics.Methods A total of 40 healthy rabbits were randomly divided into a normal group,a model group,a reinforcing-reducing needling(RRN)group,a twirling-reinforcing needling(TRN)group and a HRN group(n=8rabbits/group).The RA model with cold syndrome was established with ovalbumin and freezing.Except the normal and model groups,RRN,TRN and HRN were given at"Zusanli"(ST 36)in the corresponding acupuncture groups for 30 min,once a day for 7days.After the interventions,the pain threshold and the local skin temperature of each group were observed.The endogenous metabolites in the serum were analyzed by the gas chromatography mass spectrometry(GC-MS)technology combined with the pattern-recognition method.Results The pain threshold and the local skin temperature in the model group were lower than those in the normal group(P<0.05).The pain thresholds and the local skin temperatures in the three acupuncture groups were higher than those in the model group after interventions(P<0.05),and those in the HRN group were obviously higher than those in the RRN and TRN groups(P<0.05).The serum metabolites of leucine,valine,isoleucine,and arachidonic acid in the model group were increased compared with those in the normal group(P<0.05),and the metabolites ofα-ketoglutaric acid,citric acid,succinic acid,glucose,inositol,palmitic acid,stearic acid,lactose,d-ribose,and D-mannose were decreased(P<0.05).The above-mentioned metabolites in the three acupuncture groups were significantly reversed relevant to the model group(P<0.05),and the effect of HRN group was significantly superior to those of the RRN and TRN groups in regulatingα-ketoglutaric acid,citric acid,succinic acid,glucose,inositol,d-ribose,and D-mannose(P<0.05).Conclusion HRN for RA with cold syndrome is effective,which may be related to the specific regulation for the krebs cycle and glycometabolism.

关键词(KeyWords)： 热补针法;类风湿关节炎;代谢组学
Heat-reinforcing needling;Rheumatoid arthritis;Metabonomics

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(No.81260558);; 国家中医药管理局甘肃郑氏针法学术流派传承工作室项目(No.2305135901)

作者(Authors): 杜小正;袁博;张星华;田亮;张婷卓;李兴兰;张枫帆;王金海;
DU Xiao-zheng;YUAN Bo;ZHANG Xing-hua;TIAN Liang;ZHANG Ting-zhuo;LI Xing-lan;ZHANG Feng-fan;WANG Jin-hai;School of Acu-moxibustion and Massage,Gansu University of Chinese Medicine;Department of Acu-moxibustion,Gansu Provincial Hospital of Traditional Chinese Medicine;Department of Chinese Medicine,the Second Hospital of Lanzhou University;

DOI: 10.13702/j.1000-0607.2017.03.003

参考文献(References)：

• [1]BURSKA A,BOISSINOT M,PONCHEL F.Cytokines as biomarkers in rheumatoid arthritis[J].Mediators Inflamm,2014,2014:545493.
• [2]邢巍.略谈类风湿性关节炎的中医辨治[J].中国民族民间医药,2010,20(2):7-8.
• [3]张星华,朱博雯,杨晨光,等.热补针法治疗风寒湿阻型类风湿性关节炎的临床疗效[J].辽宁中医杂志,2015,42(5):1076-1078.
• [4]杜小正,方晓丽,东贵荣.热补针法对关节炎兔的镇痛后效应及其脑脊液中β-EP和CCK-8含量的影响[J].中医研究,2011,24(6):18-20.
• [5]杜小正,王金海,张星华,等.热补针法对风寒湿阻型类风湿关节炎患者血浆中能量代谢相关基因Atp 5O、Atp 6V1B2表达的影响[J].针刺研究,2016,41(4):303-307.
• [6]杜小正,秦晓光,方晓丽.热补针法镇痛后效应及其对关节局部组织β-内啡肽和前列腺素E 2的影响[J].针刺研究,2009,34(5):319-323.
• [7]杜小正,王金海,秦晓光,等.热补针法对RA模型家兔关节滑膜LDH、ATPase活性的影响[J].中医研究,2016,29(1):58-62.
• [8]NICHOLSON J K,LINDON J C,HOLMES E.‘Metabonomics’:understanding the metabolic responses of living systems to pathophysiological stimuli via multivariate statistical analysis of biological NMR spectroscopic data[J].Xenobiotica,1999,29(11):1181-1189.
• [9]NICHOLSON J K,LINDON J C.Systems biology:metabonomics[J].Nature,2008,455(7216):1054-1056.
• [10]孙皓,刘薇,曾建国,等.GC-MS分析不同产地香荚兰商品荚乙醇提取物化学成分差异及其香草醛的测定[J].中草药,2013,44(8):955-959.
• [11]TOMINAGA K,ALSTERGREN P,KURITA H,et al.Clinical course of an antigen induced arthritis model in the rabbit temporomandibular joint[J].J Oral Pathol Med,1999,28(6):268-273.
• [12]吴垦莉,张珊珊.寒凝血瘀证动物模型的研制[J].中国中医基础医学杂志,1996,2(2):49-51.
• [13]李忠仁.实验针灸学[M].北京:中国中医药出版社,2007:242-247.
• [14]陆寿康.刺法灸法学[M].北京:中国中医药出版社,2003:92-95.
• [15]郑魁山,郑俊江,郑俊鹏,等.郑氏针灸全集[M].北京:人民卫生出版社,2000:289-290.
• [16]GU Y,LU C,ZHA Q,et al.Plasma metabonomics study of rheumatoid arthritis and its Chinese medicine subtypes by using liquid chromatography and gas chromatography coupled with mass spectrometry[J].Mol Bio Syst,2012,8(5):1535-1543.
• [17]ZHOU J,CHEN J,HU C,et al.Exploration of the serum metabolite signature in patients with rheumatoid arthritis using gas chromatography-mass spectrometry[J].J Pharm Biomed Anal,2016,127:60-67.
• [18]PRIORI R,CASADEI L,VALERIO M,et al.1 H-NMRbased metabolomic study for identifying serum profiles associated with the response to etanercept in patients with rheumatoid arthritis[J].PLoS One,2015,10(11):e 0138537.
• [19]WANG Y,GUO X,XIE J,et al.A GC-MS based metabonomics study of rheumatoid arthritis and the interventional effects of the Simiaowan in rats[J].Molecules,2015,20(12):21364-21372.
• [20]YE T S,DU Z H,LI Z H,et al.Repeated electroacupuncture persistently elevates adenosine and ameliorates collageninduced arthritis in rats[J].Evid Based Complement Alternat Med,2016,2016:3632168.
• [21]张伟,陈明人,胡锦玉,等.不同发泡灸量的督脉铺灸治疗类风湿关节炎的临床疗效观察[J].针刺研究,2016,41(4):327-333.
• [22]SEMEGA Y,MBOW C,SUH S J,et al.The effect of BCAA supplementation upon the immune response of triathletes[J].Med Sci Sports Exerc,2000,32(7):1214-1219.
• [23]BASSIT R A,SAWADA L A,BACURAU R F P,et al.Branched-chain amino acid supplementation and the immune response of long-distance athletes[J].Nutrition,2002,18(5):376-379.
• [24]WANG M,WANG B,MA Z,et al.Detection of the novel IL-1family cytokines by QAH-IL 1F-1assay in rheumatoid arthritis[J].Cell Mol Biol(Noisy-le-grand),2016,62(4):31-34.
• [25]KOKKONEN H,SDERSTRM I,ROCKLV J,et al.Up-regulation of cytokines and chemokines predates the onset of rheumatoid arthritis[J].Arthritis Rheum,2010,62(2):383-391.
• [26]洪寿海,丁沙沙,张阔,等.基于细胞因子的针刺治疗类风湿关节炎的镇痛、抗炎机制研究进展[J].针刺研究,2016,41(5):469-473.
• [27]张瑞,郭丽红,殷媛,等.电针对胶原诱导性关节炎大鼠血清中炎性细胞因子含量的影响[J].针刺研究,2016,41(1):51-54.
• [28]郭丽红,刘长信,田贵花,等.电针对类风湿性关节炎大鼠血清及膝关节白介素-17和白介素-23的影响[J].针刺研究,2015,40(2):141-145.
• [29]GENNARO A D,HAEGGSTRM J Z.The leukotrienes:immune-modulating lipid mediators of disease[J].Adv Immunol,2012,116:51-92.
• [30]DI G A,HAEGGSTRM J Z.Targeting leukotriene B 4in inflammation[J].Expert Opin Ther Targets,2014,18(1):79-93.
• [31]YOUSEFI B,JADIDI-NIARAGH F,AZIZI G,et al.The role of leukotrienes in immunopathogenesis of rheumatoid arthritis[J].Mod Rheumatol,2014,24(2):225-235.
• [32]WANG H,YE J.Regulation of energy balance by inflammation:common theme in physiology and pathology[J].Rev Endocr Metab Disord,2015,16(1):47-54.
• [33]SIVAKUMAR B,AKHAVANI M A,WINLOVE C P,et al.Synovial hypoxia as a cause of tendon rupture in rheumatoid arthritis[J].J Hand Surg Am,2008,33(1):49-58.
• [34]TRIPMACHER R,GABER T,DZIURLA R,et al.Human CD 4(+)T cells maintain specific functions even under conditions of extremely restricted ATP production[J].Eur J Immunol,2008,38(6):1631-1642.